Affinity purification followed by mass spectrometry has become the technique of choice to identify binding partners in biochemical complexes isolated from a physiologic cellular context. In this report we detail our protocol for tandem affinity purification (TAP) primarily based on the use of the FLAG and HA peptide epitopes, with a particular emphasis on factors affecting yield and specificity, as well as steps to implement an automated version of the TAP procedure. © 2019 by John Wiley & Sons, Inc.
Keyphrases
- mass spectrometry
- capillary electrophoresis
- liquid chromatography
- gas chromatography
- high performance liquid chromatography
- high resolution
- randomized controlled trial
- binding protein
- multiple sclerosis
- ms ms
- amino acid
- dna binding
- protein protein
- tandem mass spectrometry
- small molecule
- simultaneous determination
- hepatitis c virus